Building Better Antioxidants: Virtual Screening, Synthesis, and Characterization of Multifunctional Small Molecules Combining Nrf2 Pathway Activation and Direct Antioxidant Activity

项目摘要/摘要 失控的氧化应激导致神经退行性疾病和眼部疾病的发生 目前还没有有效的治疗干预措施。调查人员已经开发出了小 在生物检测中通过两种机制表现出显著抗氧化活性的分子：直接 与活性氧物种的反应性和Nrf2途径的催化激活。调查人员将 结合虚拟筛选、合理设计、合成化学和分子生物学方法 生产增强的抗氧化剂小分子，并了解小分子如何激活催化剂 抗氧化途径。 从这个项目中产生的新分子有望提供清除自由基的活性，代谢 稳定性和渗透性。我们提出了四种改进这些特性的机制 与母体分子相比。将使用虚拟筛选来确定这些项目的哪种组合 方法将产生最有希望的目标。在合成这些靶点之后，细胞模型 神经退行性疾病和眼部疾病(如白内障)将被用来测试对氧化应激的保护。 然后对表现出强大的抗氧化活性的分子进行代谢稳定性和血脑筛选。 屏障渗透性。探索还将集中在揭示我们的 小分子通过催化激活生物体内的天然抗氧化剂途径进行保护(NRF2)。细胞 暴露于ROS的模型的裂解产物将被评估Nrf2及其下游基因，包括NQO1， 例如，HO-1、SOD、GPX和TrX。本文还探讨了Nrf2与炎症的关系。 这种方法将确定每个抗氧化增强策略提供的保护路径(S)和 确定有待进一步探索的铅分子，并继续进行未来涉及动物毒理学的工作， 清除和活动评估。 总之，使用化学分析和生物学研究数据的比较方法将使 研究人员识别提供所需特征的分子和部分，以作为 治疗氧化应激引起的神经退行性和眼部疾病。这项提议需要 通过使用计算化学和合成化学相结合来针对氧化应激的独特方法 将不同的反应性积木转化为小分子，旨在通过靶向发挥作用 神经退行性变和眼睛疾病的分子特征大于 各个部件。

PROJECT SUMMARY/ABSTRACT Uncontrolled oxidative stress contributes to the development of neurodegenerative and ocular diseases for which there are yet to be effective therapeutic interventions. The investigators have developed small molecules that exhibit significant antioxidant reactivity in biological assays through two mechanisms: direct reactivity with reactive oxygen species and catalytic activation of the Nrf2 pathway. The investigators will incorporate virtual screening, rational design, synthetic chemistry, and molecular biology methods to produce enhanced antioxidant small molecules and understand how the small molecules activate catalytic antioxidant pathways. New molecules produced from this project are expected to provide radical scavenging activity, metabolic stability, and permeability. We propose four mechanisms of achieving improvement in these features compared to parent molecules. Virtual screening will be used to identify what combination of these approaches will yield the most promising targets. Following synthesis of these targets, cellular models for neurodegenerative and ocular disease (e.g. cataracts) will be used to test for protection from oxidative stress. Molecules showing potent antioxidant activity will then be screened for metabolic stability and blood brain barrier permeability. Explorations will also be focused on uncovering the mechanism(s) through which our small molecules protect through catalytic activation of innate antioxidant pathways in biology (Nrf2). Cell lysates from models exposed to ROS will be evaluated for Nrf2 and its downstream genes including NQO1, HO-1, SOD, GPX, and Trx, for example. The relationship between Nrf2 and inflammation is also explored. This approach will identify the path(s) of protection each strategy of antioxidant enhancement provides and identify lead molecules to be explored further and proceed to future work involving animal toxicology, clearance, and activity assessment. Altogether, a comparative approach that uses data from chemical assays and biological studies will allow the investigators to identify molecules and moieties that provide the characteristics needed to serve as a therapeutic for neurodegenerative and ocular disorders arising from oxidative stress. This proposal takes a unique approach to targeting oxidative stress by using computational and synthetic chemistry to combine different reactive building-blocks into small molecules designed to have activity through targeting molecular features of neurodegeneration and diseases of the eye in a manner that is greater than the sum of the individual parts.